Green Technology Approach for Reinforcement of Calcium Chloride Cured Sodium Alginate Films by Isolated Bacteria from Palm Oil Mill Effluent (POME)

Round 1

Reviewer 1 Report

This paper deals with the benefits of adding bacteria to the Na alginate films. The authors have discussed the process of adding bacteria and the evaluation of the mechanical properties of the films. The experimental findings support the conclusions drawn. There are, however, a few concerns listed below that need to be addressed before this paper is accepted for the publication.

1. Rewrite Line 75, as it is hard to understand.

2. Line 105: Mention the source of purchase

3. Line 143: replace "Constant Variable" with "control sample" or "base sample".

4. Line 198: Check the formatting.

5. Reference 16: Check the formatting.

Section 3.6: Please explain how the bacteria prevented/reduced water absorption. The authors mention that the presence of bacteria absorbs water due to osmosis. Referring to Figure 4, the bacteria is present sparingly on the surface of the sample. I wonder how could a small number of bacteria change water absorption behavior.

6. It will be helpful if the authors can add more explanation to the changes in the properties of the films due to the addition of bacteria.

7. Adding graphs showing the changes in the properties with different amounts of bacteria will help understand this work better.

 

Author Response

Reviewer’s Comments	Response
1. Rewrite Line 75, as it is hard to understand.	Line 75 has been revised into two different sentences and easier to understand.
2. Line 105: Mention the source of purchase	The source of purchase of the chemicals has been mentioned.
3. Line 143: replace "Constant Variable" with "control sample" or "base sample".	"Constant Variable" has been replaced with with "control sample".
4. Line 198: Check the formatting.	The formatting of Line 198 has been revised accordingly.
5. Reference 16: Check the formatting.	The formatting of Reference 16 has been revised accordingly.
Section 3.6: Please explain how the bacteria prevented/reduced water absorption. The authors mention that the presence of bacteria absorbs water due to osmosis. Referring to Figure 4, the bacteria is present sparingly on the surface of the sample. I wonder how could a small number of bacteria change water absorption behavior.	Size of bacteria have been proven by many previous researchers are micro size and it has capability to sealed crack, pores or voids in the micro size too. Once the cracks, pores or voids are sealed, reduction in water ingress is observed. Obviously, bacterial activity or cell can seal the pores, voids and micro cracks, where other sealants are unable to work through (Chahal et al., 2012). (Added in line 311-314)   Other reason also has been elaborate in the line 300-310
6. It will be helpful if the authors can add more explanation to the changes in the properties of the films due to the addition of bacteria.	Elaborated in the section 3.7 and 3.8
7. Adding graphs showing the changes in the properties with different amounts of bacteria will help understand this work better.	Added in the figures 4,5 and 6

Reviewer 1

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript entitled “Green Technology Approach for Reinforcement of  Calcium Chloride Cured Sodium Alginate Films by Sub-micron Bacillus Bacteria  “ written by Briant Ho Kang Xian et all presents the use of the sub-micron bacillus bacteria as fillers to reinforce the calcium chloride cured sodium alginate film. The interaction between the bacteria fillers and the component of the composite  film was determined by using SEM analysis. The results revealed that the composite films reinforced with 0.3 g and 0.4g  of bacteria appeared to have less curling on the surface of the film. The conclusion were bacteria fillers do not possess the  ability  to  enhance  the  elongation. Generally it was concluded that the bacteria biomass has the potential to be used as fillers to reinforce calcium chloride cured sodium alginate film

 

Comments

1) The authors claim that elongation was achieved with 0.3 g and 0.4g  bacteria, but they did not present data for other values eg 0.5. maybe the trend of increasing elongating is still growing up. Could author comments on this.

2)  Could authors presents more SEM images of the films . The modification of the figure 2 with comparison of SEM  images with different addition of bacteria would improve quality of the manuscript.

3) The main manuscript presents 4 figures which easily could be contended into two figures.  First one with images with 10 micrometres scale and second figures with both images with 50 micrometres scale. Figure 7 from the supplementary materials should be included into main manuscript. The figures 8,9,10 from the supplementary materials should be connected into one big table and moved into main manuscript. Right now,  the manuscript looks rather poor having only three single SEM images. Moving figures from supplementary material into main text will increase quality of the manuscript.

4) There are some editorial mistakes, formatting , missing spaces are easily find thought whole manuscript. Please correct it  eg  Line 198,

Author Response

Reviewer 2

Reviewer’s Comments	Response
1. Line 25-26 sentence is not clear, please revise	Line 25-26 have been revised accordingly to make the sentences clearer to be understood.
2. Line 49-50 contains the wrong information.	Line 49-50 have been revised and amended accordingly. The last sentence of Line 50 has been removed.
3. Line 73-74 not necessary.	The sentence of Line 73-74 has been removed.
4. Materials and methods section, please mention the sub-topics such as materials; bacteria isolation, bacteria identification; mechanical properties measurement etc.	Done
5. Line106, need purity and company information of chemicals.	Done
6. The authors autoclave the bacteria before incorporation into hydrogel. My concern is bacteria can be destroyed because of that high temperature during autoclaving. If damaged, the purpose of this study is nothing. If not destroyed please show the sem images of bacteria only before and after autoclaving.	Actually the sentences need to rephrase. It corrected under section methodology, sub section  Preparation Mass Volume of Bacteria
7. mL and ml (line 142), make consistent writing throughout MS	All the ‘ml’ in the manuscript have been changed to ‘mL’.
8. 156, crosslinking time is very short. What happens if crosslinked more time?
9. In mechanical study, if properties were not calculated manually, no need to write the formulae. The UTM machine gives the data automatically.	The formulae have been removed accordingly.
10. Line 198, correct	Line 198 has been corrected accordingly.
11. Lines 202-203 not necessary	Line 202-203 has been removed accordingly.
12. All Supplementary files should adjust in the main MS.	Done
13. Section 3.2 contains the general information which is not necessary.	The general information in Section 3.2 has been revised and removed accordingly.
14. Need biochemical analysis of the main bacterium that incorporated in film. Only gram staining is not enough.	Actually, this research  focus on isolated the bacteria and morphology test (shape and gram staining) purpose to ensure that colonies of bacteria is not come from the same species.  We not do any detail characterization and identification of bacteria.
15. Too many decimals are presented in tables and texts. The results cannot be guaranteed to be such accuracy.	Amended.
16. 4 contains too many unnecessary information. Please revise. Please write scientifically.	The conclusion has been revised accordingly. Unnecessary information has been removed and revised.
17. Figure 1-4 should be merged in one figure so make to compare easier.	Done

 

 

 

Author Response File: Author Response.docx

Reviewer 3 Report

This paper talks about the reinforcement of alginate hydrogel film by the bacteria. The manuscript is poorly prepared, contained many unnecessary information.  Ms contains 40% junk information so please shorten. Needs extensive English correction. Few important concerns are as follows

1. Line 25-26 sentence is not clear, please revise
2. Line 49-50 contains the wrong information.
3. Line 73-74 not necessary.
4. Materials and methods section, please mention the sub-topics such as materials; bacteria isolation, bacteria identification; mechanical properties measurement etc.
5. Line106, need purity and company information of chemicals.
6. The authors autoclave the bacteria before incorporation into hydrogel. My concern is bacteria can be destroyed because of that high temperature during autoclaving. If damaged, the purpose of this study is nothing. If not destroyed please show the sem images of bacteria only before and after autoclaving.
7. mL and ml (line 142), make consistent writing throughout MS
8. 156, crosslinking time is very short. What happens if crosslinked more time?
9. In mechanical study, if properties were not calculated manually, no need to write the formulae. The UTM machine gives the data automatically.
10. Line 198, correct
11. Lines 202-203 not necessary
12. All Supplementary files should adjust in the main MS.
13. Section 3.2 contains the general information which is not necessary.
14. Need biochemical analysis of the main bacterium that incorporated in film. Only gram staining is not enough.
15. Too many decimals are presented in tables and texts. The results can not be guaranteed to be such accuracy.
16. 4 contains too many unnecessary informations. Please revise. Please write scientifically.
17. Figure 1-4 should be merged in one figure so make to compare easier.

 

 

Author Response

no have 3 reviewers

Round 2

Reviewer 1 Report

All my comments have been addressed. This paper is acceptable in the present form. 

Reviewer 2 Report

The manuscript entitled “Green Technology Approach for Reinforcement of Calcium Chloride Cured Sodium Alginate Films by Isolated Bacteria from Palm Oil Mill Effluent (POME) “ written by Briant Ho Kang Xian et all presents the use of the sub-micron bacillus bacteria as fillers to reinforce the calcium chloride cured sodium alginate film.  The authors presents morphological and mechanical properties of composite films. The cross-sectional  morphologies  of  the  films   were  observed  by  using  scanning  electron.  The conclusion is bacteria fillers do not possess the ability to enhance the elongation properties because only about 0.6% of overall increment observed, which was considered insignificant. It was concluded that the bacteria biomass has the potential to be used as fillers to reinforce calcium chloride cured sodium alginate film.

 

Comments

1) The Figure one needs to be improved. The Quality of the images it not very good in panel a ,maybe author have images with better resolution. The scale bar is not visible. Authors could make comments in the figure caption that scale bar is 20 micrometres.

2) There are some editorial mistakes, formatting , missing spaces are easily find thought whole manuscript. Please correct it  eg  Line 399 there is  “a)Cross-sectional” the space between "a) Cross-sectional” is missing

Reviewer 3 Report

the authors improve the MS satisfactorily.